Journal: Oncotarget

Article Title: Branched multipeptide immunotherapy for glioblastoma using human leukocyte antigen-A*0201-restricted cytotoxic T-lymphocyte epitopes from ERBB2, BIRC5 and CD99

doi: 10.18632/oncotarget.10495

Figure Lengend Snippet: a. ERBB2 peptides, b. BIRC5 peptides, c. CD99 peptides. ERBB2 369 (KIFGSLAFL), BIRC5 96methionine (LMLGEFLKL) and CD99 9 (LLLFGLLGV) were selected as HLA-A*0201-specific peptides with high scores in the peptide-T2 binding assay. The concentration of each peptide was 20 μg/mL. The values represent the mean MFI ± SE of three separate experiments.

Article Snippet: The T2 cell line, a human B and T cell hybrid expressing HLA-A2 molecules, was obtained from the Korean Cell Line Bank, Seoul, Korea.

Techniques: Binding Assay, Concentration Assay

Journal: Oncotarget

Article Title: Branched multipeptide immunotherapy for glioblastoma using human leukocyte antigen-A*0201-restricted cytotoxic T-lymphocyte epitopes from ERBB2, BIRC5 and CD99

doi: 10.18632/oncotarget.10495

Figure Lengend Snippet: a. Binding affinity: The multipeptide cocktail dose-dependently increased the HLA-A02 mean MFI in T2 cells. In cells treated with the branched multipeptide, the mean MFI increased until the total peptide concentration reached 22.5 μg/mL (7.5 μg/mL/peptide), which was selected as the optimal peptide concentration. b. Binding stability: While the stability of the branched multipeptides was maintained up to 24 hours post-BFA treatment, the stability of the multipeptide cocktail was only maintained up to 8 hours post-BFA treatment. The values represent the mean MFI ± SE of three separate experiments.

Article Snippet: The T2 cell line, a human B and T cell hybrid expressing HLA-A2 molecules, was obtained from the Korean Cell Line Bank, Seoul, Korea.

Techniques: Binding Assay, Concentration Assay

Journal: The Journal of Experimental Medicine

Article Title: Efficient Generation of a Hepatitis B Virus Cytotoxic T Lymphocyte Epitope Requires the Structural Features of Immunoproteasomes

doi:

Figure Lengend Snippet: Time-dependent generation of a potential precursor peptide HBcAg141-162 and of HBcAg141-151. HBcAg131-162 was digested for 0.5, 1, 2, 4, 8, and 24 h with 20S proteasome isolated from T2 cells and T2 LMP2+7 transfectant cells. Cleavage products were separated by HPLC as described in Materials and Methods but analyzed on-line by an ion trap mass spectrometer (ThermoQuest) equipped with an electrospray ion source, which results in increased sensitivity. Time course of the potential precursor peptide HBcAg141-162, which is generated by a single cleavage after Leu140 (A), and of the antigenic HBcAg141-151 (B) is depicted. Plotted values were confirmed by repeated analysis of the same digests.

Article Snippet: C4 (BALB/c) mouse fibroblast cells were cultured in Iscove's MEM (Biochrom), and the human cervical carcinoma cell line HeLa, the human lymphoblastoid cell line T2 , and T2 transfectant cell lines were cultured in RPMI (Biochrom).

Techniques: Isolation, Transfection, Mass Spectrometry, Generated

Journal: The Journal of Experimental Medicine

Article Title: Efficient Generation of a Hepatitis B Virus Cytotoxic T Lymphocyte Epitope Requires the Structural Features of Immunoproteasomes

doi:

Figure Lengend Snippet: Incorporation of the LMP2 and LMP7 subunits in 20S proteasomes of T2 transfectant lines. 500 ng (A) or 150 ng (B) of purified 20S proteasomes was separated by SDS-PAGE and probed with mouse LMP2–specific (A) or mouse LMP7–specific (B) polyclonal rabbit antisera. Molecular mass markers (kD) are shown at left.

Article Snippet: C4 (BALB/c) mouse fibroblast cells were cultured in Iscove's MEM (Biochrom), and the human cervical carcinoma cell line HeLa, the human lymphoblastoid cell line T2 , and T2 transfectant cell lines were cultured in RPMI (Biochrom).

Techniques: Transfection, Purification, SDS Page

Journal: The Journal of Experimental Medicine

Article Title: Efficient Generation of a Hepatitis B Virus Cytotoxic T Lymphocyte Epitope Requires the Structural Features of Immunoproteasomes

doi:

Figure Lengend Snippet: Kinetic analysis of the production of HBcAg141-151 and of related peptide fragments. HBcAg131-162 was digested for 4, 8, or 24 h with 20S proteasomes isolated from T2 LMP2+7 (A) and T2 LMP2+7 T1A transfectant cells (B) or mock-digested (C). Cleavage products were analyzed as described in Materials and Methods. Relative abundances of 11 selected peptides are plotted. Shown data were reproduced in a second, independent experiment.

Article Snippet: C4 (BALB/c) mouse fibroblast cells were cultured in Iscove's MEM (Biochrom), and the human cervical carcinoma cell line HeLa, the human lymphoblastoid cell line T2 , and T2 transfectant cell lines were cultured in RPMI (Biochrom).

Techniques: Isolation, Transfection